Abstract
We investigated the fruits of Rubus crataegifolius Bge, a plant which has been traditionally used in Korea in phytotherapy, to describe antioxidant materials from plant sources. R. crataegifolius fruits were extracted with methanol and further fractionated into n-hexane, diethyl ether, and ethyl acetate. The antioxidant activity of each fraction and the residue was assessed using a 1,1-diphenyl-2-picrylhydrazyl (DPPH), $H_2O_2$ radical scavenging method, and their cytotoxicity on human primary kerationcyte (HK) was determined by an MTS assay. The R. crataegifolius fruit methanol extract showed strong antioxidant activity (75.04%, 50%) compared with vitamin C (79.9%, 54.1%) by the DPPH, and $H_2O_2$ method, respectively. The measured activity from the subsequent extracts of the methanol extract were 20.3% for n-hexane fraction (HF), 68.8% for diethyl ether fraction (DF), 67.1% for ethyl acetate fraction (EF), and 67.1% for the residue fraction (RE) by DPPH and 2.2% for HF, 1.6% for DF, 10% for EF, and 50% for the RE by $H_2O_2$ assay. An oxidative stress model of HK was established under a suitable concentration (1 mM). The cell viability of the RE treated group increased and the percentage of apoptotic cells decreased at concentrations of 0.005-0.02% RE compared with the $H_2O_2$ treated group. Fruit extracts of the medicinal plant R. crataegifolius showed potent antioxidant activity and the ability to relieve cell damage from $H_2O_2$ induced injury to HK.
식물자원으로부터 항산화 물질을 확보 하기위해 한국 등에서 전통적으로 phytotherapy로 이용되어 온 Rubus crataegifolius Bge. 열매를 조사하였다. R. crataegifolius의 열매를 메탄올로 추출하였고 순차적으로 n-hexane, diethyl ether, and ethyl acetate로 분획화하였다. 각 분획물의 항산화 활성은 DPPH와 $H_2O_2$에 대항 인간 primary 세포인 keratinocyte (HK)를 이용하여 세포 독성 및 효능을 검증하였다. R. crataegifolius 열매 추출물은 비타민 C와 비슷한 강력한 DPPH (75.04%, 50%)와 $H_2O_2$ (79.9%, 54.1%) 소거능을 보였다. 분획물의 DPPP에 대한 소거능을 측정하였는데 n-hexane fraction (HF)은 20.3%, diethyl ether fraction (DF)은 68.8%, ethyl acetate fraction (EF)는 67.1% 그리고 residue fraction (RE)은 67.1%의 소거능을 보였으며 $H_2O_2$에 대해서는 2.2%, 1.6%, 10%, 그리고 50%로 각각 나타내었다. H2O2에 대한 HK의 세포 보호능을 확인하기 위해 산화적 스트레스 모델을 확립하였고(1 mM) 0.005-0.02%의 RE 분획물에서 $H_2O_2$에 대한 보호능을 발휘하였다. 따라서, R. crataegifolius의 열매 추출물은 $H_2O_2$유발 상처에 대하여 HK세포의 보호능을 가지며 강력한 항산화 활성을 가지고 있다.
